Yanjian Wan

Quantification of microRNA by gold nanoparticle probes.

MicroRNAs are important posttranscriptional regulators of gene expression in animals and plants. A sensitive and specific detection method is urgently needed for intensive studies on differential expression and regulatory roles of microRNA. Here we present a simple and reliable method for the quantification of microRNA. The hybridization products of target microRNA with capture probe and gold nanoparticle probe are immobilized onto the surface of a streptavidin-coated microplate, and the signal is amplified by silver enhancement. Distribution of miR-122a/miR-128 in mouse brain and liver tissue was detected by this method, and synthetic miRNA122a was quantified. This method allowed a lower detect limit of 10 fM with a linear dynamic range from 10 pM to 10 fM and a high specificity to discriminate one single oligonucleotide mismatch of the target microRNA.

Prenatal PFOS exposure induces oxidative stress and apoptosis in the lung of rat off-spring

Perfluorooctane sulfonate (PFOS) could induce neonatal pulmonary injuries in rodents. The aim of this study was to investigate the underlying mode of action. Pregnant rats were dosed orally with PFOS (0, 0.1 and 2.0mg/kgd) from gestation days (GD) 1 to 21. Lung samples from postnatal day (PND) 0 and 21 pups were analyzed for the toxic effects of PFOS. The results showed that maternal exposure to 2.0mg/kgd PFOS caused severe histopathological changes along with marked oxidative injuries and cell apoptosis in offspring lungs; at the same time, the ratio of Bax to Bcl-2, release of cytochrome c (Cyt c) from mitochondria to cytoplasm, expressions of Fas and Fas-L, and activities of caspase-3, -8 and -9 were up-regulated correspondingly. The results indicate that oxidative stress and both intrinsic and extrinsic cell death pathways were involved in prenatal PFOS exposure-induced injuries in postnatal lungs.

Electrochemical biosensor for estrogenic substance using lipid bilayers modified by Au nanoparticles.

There is a growing demand for new technologies that are capable of screening the wide variety of xenoestrogens in environment. Here, a nanostructure electrochemical biosensor was developed to directly detect and screen estrogenic substances based on estrogen receptor (ER) binding without the use of radio- or enzyme-labeled compounds. The biosensor was fabricated by immobilization of ERs in supported bilayer lipid membrane (s-BLM) modified with Au nanoparticles, and the properties of the modified electrodes were characterized by cyclic voltammetry and impedance spectroscopy. The results indicated that the biosensor was able to detect the natural estrogen 17beta-estradiol with an acceptable linear correlation ranging from 5 to 150 ng/L and a detection limit of 1 ng/L. The biosensor could also detect other known xenoestrogens such as bisphenol A and 4-nonylphenol with satisfied sensitivity and quantitative results. The biosensor showed good reliability and repeatability, and the Au nanoparticles greatly enhanced the sensitivity and stability of the biosensor. Moreover, estrogenic activity of water samples determined by this biosensor was in good agreement with that determined by MCF-7 cell proliferation assay.

Early-Life Exposure to Bisphenol A Induces Liver Injury in Rats Involvement of Mitochondria-Mediated Apoptosis

Exposure to bisphenol A (BPA), a monomer widely used to manufacture polycarbonate plastics, has been reported to be associated with abnormalities of liver function and hepatic damage. However, the molecular mechanism under the pathogenesis of hepatic injury is unclear. In this study, the effect of perinatal exposure to BPA at the reference dose of 50 µg/kg/day on the apoptotic index in the liver of rat offspring was investigated. Increased levels of ALT and enhanced cell apoptosis were observed in the liver of rat offspring at 15 and 21 weeks, and significantly increased activity of caspase-3 and caspase-9 and elevated levels of cytochrome c were also confirmed. In addition, significant change in the expression levels of Bcl-2 and Bax were found in BPA-treated offspring at 21 weeks. For in vitro experiments, liver mitochondria were isolated from neonatal rats and were treated with BPA. BPA treatment led to a significant increase in mitochondrial permeability transition. Moreover, the supernatant from BPA-treated mitochondria significantly increased apoptotic changes in nuclei isolated from liver tissue. In conclusion, the study demonstrates that BPA induces mitochondria-mediated apoptosis in hepatic cells, which may contribute to long-term hepatotoxicity induced by early-life exposure to BPA.

Alterations in tumor biomarker GSTP gene methylation patterns induced by prenatal exposure to PFOS

The adverse environmental exposure in early life may have adverse effects on animals through epigenetic aspects. The current study examined the possibility of early epigenetic alteration in PFOS-exposed rat liver. Pregnant Sprague-Dawley (SD) rats were exposed to perfluorooctane sulfonate (PFOS) at doses of 0.1, 0.6 and 2.0 mg/kg/d and 0.05% Tween 80 as control by gavage from gestation days 2 to 21. The dams were allowed to give birth and liver samples from weaned (postnatal day 21) offspring rats were analyzed for PFOS content, relative liver weight, global DNA methylation, methylation of LINE-1 regulatory region, tumor suppressor gene glutathione S-transferase pi (GSTP) and p16 promoter methylation level, as well as related genes expression level. In PFOS-exposed weaned rats, compared to the control, global DNA methylation and methylation of LINE-1 regulatory region decreased significantly only in the 2.0 mg/kg/d group. Up to 30% of critical CpG sites (+79, 81 and 84) in GSTP promoter region were methylated in the livers of PFOS-treated rats, while p16 promoter methylation was not affected. In addition, the up-regulated expression of GSTP was observed and this increase was associated with its main pathway of transcription regulation: Keap1-Nrf2/MafK. Thus, early-induced changes in critical cytosines within the GSTP gene promoter region may be a biomarker of hepatic PFOS burden, though their direct role in PFOS-induced hepatotoxicity, including its potential carcinogenic action, needs further research.

MicroRNA expression changes during zebrafish development induced by perfluorooctane sulfonate

Perfluorooctane sulfonate (PFOS), a kind of widely distributed environmentally organic compound, has been found to cause developmental toxicity. Although microRNAs (miRNAs) play an important role in many metabolic tasks, whether and how they are involved in the process of PFOS‐induced toxicity is largely unknown. To address this problem, PFOS‐induced changes in miRNAs and target gene expression in zebrafish embryos, and the potential mechanism of PFOS‐induced toxic action were studied in this research. Zebrafish embryos were exposed to 1 µg ml−1 PFOS or DMSO control from 6 h post‐fertilization (hpf) to 24 or 120 hpf. Subsequently, RNA was isolated from the embryo pool and the expression profiles of 219 known zebrafish miRNAs were analyzed using microarray. Finally, quantitative real‐time polymerase chain reaction was used to validate several miRNAs expression of microarray data. The analysis revealed that PFOS exposure induced significant changes in miRNA expression profiles. A total of 39 and 81 miRNAs showed significantly altered expression patterns after PFOS exposure 24 and 120 hpf. Of the changed miRNAs, 20 were significantly up‐regulated and 19 were significantly down‐regulated (p < 0.01) at 24 hpf, whereas 41 were significantly up‐regulated and 40 were significantly down‐regulated (p < 0.01) at 120 hpf. These miRNAs were involved in development, apoptosis and cell signal pathway, cell cycle progression and proliferation, oncogenesis, adipose metabolism and hormone secretion, whereas there is still little functional information available for 32 miRNAs. Our results demonstrate that PFOS exposure alters the expression of a suite of miRNAs and may induce developmental toxicity. Copyright

Lead exposure in pheochromocytoma cells induces persistent changes in amyloid precursor protein gene methylation patterns.

It has been suggested that lead (Pb) exposure in early life may increase amyloid precursor protein (APP) expression and promote the pathogenesis of Alzheimers disease in old age. The current study examined whether the DNA methylation patterns of APP gene in rat pheochromocytoma (PC12) cells changed after Pb acetate exposure. Undifferentiated PC12 cells were exposed to three doses of Pb acetate (50, 250, and 500 nM) and one control for 2 days or 1 week. The methylation patterns of APP promoter and global DNA methylation were analyzed. The DNA methyltransferase 1 (DNMT1) expression and the level of amyloid β peptide (Aβ) were also investigated. The results showed that the exposure of the three concentrations of Pb acetate could make the APP promoter hypomethylated. The global DNA methylation level and the expression of DNMT1 were changed in the 500 nM group after 2 days exposure and in the 250 and 500 nM group after 7 days exposure. Thus, Pb may exert neurotoxic effects through mechanisms that alter the global and promoter methylation patterns of APP gene.

Maternal urinary bisphenol A levels and infant low birth weight: A nested case–control study of the Health Baby Cohort in China

BACKGROUND Exposure to bisphenol A (BPA), a known endocrine disruptor, has been demonstrated to affect fetal development in animal studies, but findings in human studies have been inconsistent. OBJECTIVES We investigated whether maternal exposure to BPA during pregnancy is associated with an increased risk of infant low birth weight (LBW). METHODS A total 452 mother-infant pairs (113 LBW cases and 339 matched controls) were selected from the participants enrolled in the prospective Health Baby Cohort (HBC) in Wuhan city, China, during 2012-2014. BPA concentrations were measured in maternal urine samples collected at delivery, and the information of birth outcomes was retrieved from the medical records. A conditional logistic regression was used to evaluate the relationship between urinary BPA levels and LBW. RESULTS Mothers with LBW infants had significantly higher urinary BPA levels (median: 4.70μg/L) than the control mothers (median: 2.25μg/L) (p<0.05). Increased risk of LBW was associated with higher maternal urinary levels of BPA [adjusted odds ratio (OR)=3.13 for the medium tertile, 95% confidence interval (CI): 1.21, 8.08; adjusted OR=2.49 for the highest tertile, 95% CI: 0.98, 6.36]. The association was more pronounced among female infants than among male infants, with a statistical evidence of heterogeneity in risk (p=0.03). CONCLUSIONS Prenatal exposure to higher levels of BPA may potentially increase the risk of delivering LBW infants, especially for female infants. This is the first case-control study to examine the association in China.

Maternal urinary cadmium concentrations in relation to preterm birth in the Healthy Baby Cohort Study in China

BACKGROUND Prenatal cadmium (Cd) exposure has been associated with adverse birth outcomes, but the findings of previous studies are inconsistent. The aim of this study was to evaluate the association between prenatal Cd exposure and birth outcomes. METHODS This study was conducted in 5364 pregnant women with a live singleton birth, who were recruited between September 2012 and October 2014 in the Healthy Baby Cohort (HBC) in Wuhan, China. Gestational age (in days) was estimated using both the womans last menstrual period (LMP) and ultrasound data. All the birth outcomes including birth weight and birth length were measured in the hospital within one hour after birth through standardized procedures. Cd was measured in maternal urine collected before delivery with inductively coupled plasma mass spectrometry. RESULTS The geometric mean of Cd concentration in maternal urine was 0.55 (range 0.01-2.85) μg/g creatinine. We found each ln-unit increase in Cd concentration (μg/g creatinine) in maternal urine was associated with decreased gestational age [adjusted β=-0.77day; 95% confidence interval (CI): -1.15, -0.39 for all infants; -0.77; 95% CI: -1.29, -0.25 for boys; and -0.80; 95% CI: -1.35, -0.25 for girls]. Increased likelihood of preterm birth (PTB) was associated with ln-unit increase in urinary Cd (μg/g creatinine) [adjusted odds ratio (OR)=1.78; 95% CI: 1.45, 2.19 for all infants; 1.97; 95% CI: 1.46, 2.65 for boys; and 1.67; 95% CI: 1.24, 2.25 for girls]. Maternal urinary Cd was not significantly associated with low birth weight (LBW) and small for gestational age (SGA). CONCLUSIONS Maternal exposure to Cd during pregnancy was associated with decreased gestational age and increased likelihood of PTB.

Occurrence of perchlorate in indoor dust from the United States and eleven other countries: Implications for human exposure

Perchlorate is a widespread environmental contaminant and potent thyroid hormone disrupting compound. Despite this, very little is known with regard to the occurrence of this compound in indoor dust and the exposure of humans to perchlorate through dust ingestion. In this study, 366 indoor dust samples were collected from 12 countries, the USA, Colombia, Greece, Romania, Japan, Korea, Pakistan, Kuwait, Saudi Arabia, India, Vietnam, and China, during 2010-2014. Dust samples were extracted by 1% (v/v) methylamine in water. Analyte separation was achieved by an ion exchange (AS-21) column and analysis was performed by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). The overall concentrations of perchlorate in dust were in the range of 0.02-104μg/g (geometric mean: 0.41μg/g). The indoor dust samples from China contained the highest concentrations (geometric mean: 5.38μg/g). No remarkable differences in perchlorate concentrations in dust were found among various microenvironments (i.e., car, home, office, and laboratory). The estimated median daily intake (EDI) of perchlorate for toddlers through dust ingestion in the USA, Colombia, Greece, Romania, Japan, Korea, Pakistan, Kuwait, Saudi Arabia, India, Vietnam, and China was 1.89, 0.37, 1.71, 0.74, 4.90, 7.20, 0.60, 0.80, 1.55, 0.70, 2.15, and 21.3ng/kgbodyweight (bw)/day, respectively. Although high concentrations of perchlorate were measured in some dust samples, the contribution of dust to total perchlorate intake was <5% of the total perchlorate intake in humans. This is the first multinational survey on the occurrence of perchlorate in indoor dust.